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Cell culture media

a cell culture and media technology, applied in the field of cell culture media, can solve the problems of increasing the difficulty of obtaining serum, reducing the nutritional content of the basic media, and affecting the production efficiency of the biotech drug, etc., and achieve the effect of enhancing the performance of serum-free media and reducing or eliminating the use of serum

Inactive Publication Date: 2006-04-06
MILLIPORE CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0032] The invention provides novel cell culture media compositions that include purified lipoprotein material that reduces or eliminates the use of serum or enhance the performance of serum-free media for cell culture. The invention also includes methods of culturing cells using the cell culture media compositions. The compositions and / or methods are useful in the culture of a variety of cell types, including, for example, hybridoma cells and / or cancer cells.
[0046] In another aspect of the present invention compositions are provided that are useful as a cell culture medium that serves to increase the yield of biological products, such as proteins, produced by the cells cultured in the media. In one embodiment, compositions can increase the yield of biological products at least 25%, 30%, 50%, 100%, 200% or 300%. In another embodiment, the biological products produced can be a peptide, such as a therapeutic or diagnostic peptide, polypeptide, protein, monoclonal antibody, immunoglobulin, cytokine (such as interferon), integrin, antigen, growth factor, cell cycle protein, hormone, neurotransmitter, receptor, fusion peptide, blood protein and / or chimeric protein.

Problems solved by technology

They can be more difficult, time-consuming and expensive (at least $250 million in production facility costs alone) to produce than synthetic drugs.
Producing biotech drugs is a complicated and time-consuming process.
Such media (often called “basal media”), however, are usually seriously deficient in the nutritional content required by most animal cells.
It is a relatively expensive material, and its use greatly increases the cost of cell culture.
In addition, it is difficult to obtain serum with consistent growth characteristics.
Further, the biochemical complexity of FBS can complicate the downstream processing of the proteins of interest, therefore raising the production costs.

Method used

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Examples

Experimental program
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Effect test

example 1

Method to Obtain Cholesterol-Rich Fraction From Bovine Serum

[0147] Starting material for a process according to the present invention can be maintained at a temperature of about 0° C. to about 50° C. Typically, the temperature is maintained at about 2° C. to about 15° C. A process according to the present invention can begin by subjecting the starting material to filtration. The filtration can be carried out utilizing one or more filtration steps. According to one embodiment, two filtration steps are sequentially utilized with filters having a nominal porosity of about 5 μ and about 1 μ. Any suitable filter in this range can be utilized.

[0148] If the starting material is serum, it is preferred to add a soluble salt, such as sodium citrate, to an ionic strength of about 0.25 to about 1. Other suitable salts include sodium chloride, sodium phosphate, potassium phosphate, ammonium sulfate and sodium sulfate. The addition of a soluble salt to the above concentration will increase the...

example 2

Use of EX-CYTE® to Reduce the Use of Serum

[0172] Methods

[0173] MK2.7 hybridoma cells were used. Seed inoculum was cultured in DME / F12 and FBS in spinners then adapted to less than 1% FBS by gradual reduction of FSB concentration. To begin the experiment, cells were washed in PBS and seeded at 1×102 cells / mL in each test condition. Batch cultures were sampled daily to monitor cell density and viability until culture viability was below 30%. Daily samples of culture supernatant were taken and processed to measure antibody production by ELISA.

[0174] Results

[0175] A combination of 0.5% EX-CYTE® and 2% FBS allowed for higher cell density and prolonged viability throughout the life of the culture as compared with 10% FBS (FIG. 1). The accumulative antibody level in the 0.5% EX-CYTE® and 2% FBS condition was more than double that of the 10% FBS culture on day 7 (FIG. 2). As a result, 0.5% EX-CYTE® effectively allowed the reduction of FBS from 10% to 2%.

example 3

Use of EX-CYTE® to Replace Serum

[0176] Methods

[0177] MK2.7 hybridoma cells were used. Seed inoculum was cultured in DMEM and FBS in spinners then adapted to less than 1% FBS by gradual reduction of FBS concentration. To begin the experiment, cells were washed in PBS and seeded at 1×105 cells / mL in each media condition. The test condition consisted of 0.75% EX-CYTE® 0.4% BSA, 6.7 ug / L sodium selenite. 10 mg / L insulin and 5.5 mg / L transferrin. (BSA (Serologicals Catalogue Number 81-068). Insulin (Serologicals Catalogue Number 4506), Transferrin (Serologicals Catalogue Number 4465)). Batch cultures were sampled daily to monitor cell density and viability until culture viability was below 10%. Daily samples of culture supernatant were taken to measure antibody production by ELISA.

[0178] Results

[0179] A combination of 0.75% EX-CYTE® and 0.4% BSA in DMEM constituted a complete serum-free media formulation. The temporary drop in culture viability in the test condition on days two and ...

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Abstract

The invention is a cell culture medium that can include reduced or no serum and that enhances the performance of serum-free media for cell culture. The medium supports the growth of cells for both small scale and large scale propagation of cells. The invention also includes a method of cultivating cells using the cell culture medium of the present invention.

Description

[0001] This application claims priority to U.S. Ser. No. 60 / 535,580 filed Jan. 9, 2004, and U.S. Ser. No. 60 / 568,084 filed May 4, 2004, the contents of which are herein incorporated by reference.FIELD OF THE INVENTION [0002] The invention is a cell culture medium that can include reduced or no serum and that enhances the performance of serum-free media for cell culture. The medium supports the growth of cells for both small scale and large scale propagation of cells. The invention also includes a method of cultivating cells using the cell culture medium of the present invention. BACKGROUND OF THE INVENTION [0003] Biotechnology drugs are medicines, such as therapeutic proteins (monoclonal antibodies, blood proteins and enzymes) that are produced by living organisms to fight disease. Unlike other medicines, biotech drugs are generally not produced synthetically, but are usually produced through microbial fermentation in mammalian cell culture. They can be more difficult, time-consumin...

Claims

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Application Information

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IPC IPC(8): C12P21/04C12N5/00
CPCC12N5/0031C12N2500/25C12N2500/36C12N2500/76C12N5/0037
Inventor ABITORABI, M. ABIGUERINI, MICHAEL N.TAYLOR, STEPHEN A.HERNANDEZ, GUADALUPE G.SIMONSEN, CHRISTIAN C.
Owner MILLIPORE CORP
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